De novo design of symmetric ferredoxins that shuttle electrons in vivo
De novo design of symmetric ferredoxins that shuttle electrons in vivo
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DOI：
10.1073/pnas.1905643116
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发表时间：
2019-07-16
影响因子：
11.1
通讯作者：
Falkowski, Paul G.
中科院分区：
文献类型：
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作者：
Mutter, Andrew C.;Tyryshkin, Alexei M.;Falkowski, Paul G.
细菌铁氧化还原蛋白的对称起源是在50多年前首次提出的，但到目前为止，还没有构建出具有功能的对称分子。据推测，现存的蛋白质是通过基因复制和随后的突变从它们对称的根中漂移而来的。现有铁氧化还原蛋白的系统发育分析支持N端和c端序列的独立进化，从而允许基于共识的对称4Fe-4S分子设计。所有设计都结合了两个[4Fe-4S]簇，并表现出在-405至-515 mV范围内强烈降低的中点电位。其中一种结构通过设计的大肠杆菌代谢途径有效地穿梭电子。这些发现表明，由对称核组成的铁氧化还原蛋白可以作为设计体内应用的新型电子转移载体的平台。外壳不对称增加了序列空间而不影响电子传递功能。
A symmetric origin for bacterial ferredoxins was first proposed over 50 y ago, yet, to date, no functional symmetric molecule has been constructed. It is hypothesized that extant proteins have drifted from their symmetric roots via gene duplication followed by mutations. Phylogenetic analyses of extant ferredoxins support the independent evolution of N- and C-terminal sequences, thereby allowing consensus-based design of symmetric 4Fe-4S molecules. All designs bind two [4Fe-4S] clusters and exhibit strongly reducing midpoint potentials ranging from -405 to -515 mV. One of these constructs efficiently shuttles electrons through a designed metabolic pathway in Escherichia coli. These finding establish that ferredoxins consisting of a symmetric core can be used as a platform to design novel electron transfer carriers for in vivo applications. Outer-shell asymmetry increases sequence space without compromising electron transfer functionality.